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Celsr1-3 Cadherins in PCP and Brain Development

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Celsr1-3 Cadherins in PCP and Brain Development CHAPTER SEVEN Celsr1–3 Cadherins in PCP and Brain Development Camille Boutin, André M. Goffinet1, Fadel Tissir1 Institute of Neuroscience, Developmental Neurobiology, Universite´ Catholique de Louvain, Brussels, Belgium 1Corresponding authors: Equal contribution. e-mail address: [email protected]; andre. [email protected] Contents 1. Celsr1–3 Expression Patterns 164 2. Celsr1: A Major Player in Vertebrate PCP 165 3. Celsr2 and 3 in Ciliogenesis 169 4. Celsr1–3 in Neuronal Migration 171 5. Celsr2 and Celsr3 in Brain Wiring 174 5.1 Motifs of Celsr important for their functions 176 References 179 Abstract Cadherin EGF LAG seven-pass G-type receptors 1, 2, and 3 (Celsr1–3) form a family of three atypical cadherins with multiple functions in epithelia and in the nervous system. During the past decade, evidence has accumulated for important and distinct roles of Celsr1–3 in planar cell polarity (PCP) and brain development and maintenance. Although the role of Celsr in PCP is conserved from flies to mammals, other functions may be more distantly related, with Celsr working only with one or a subset of the classical PCP partners. Here, we review the literature on Celsr in PCP and neural devel- opment, point to several remaining questions, and consider future challenges and possible research trends. Celsr1–3 genes encode atypical cadherins of more than 3000 amino acids ( Fig. 7.1). Their large ectodomain is composed of nine N-terminal cadherin repeats (typical cadherins have five repeats), six epidermal growth factor (EGF)-like domains, two laminin G repeats, one hormone receptor motif (HRM), and a G-protein-coupled receptor proteolytic site (GPS). This is followed by seven transmembrane domains (classic cadherins are single-pass type I proteins) and a cytoplasmic tail (CT) that varies in size and, in contrast to the ectodomain, is poorly conserved among the three members. Celsr1–3 belong to the “adhesion receptor” family of G-protein-coupled receptors (GPCRs), also referred to as long N-terminal group B (LNB) GPCRs. # Current Topics in Developmental Biology, Volume 101 2012 Elsevier Inc. 161 ISSN 0070-2153 All rights reserved. http://dx.doi.org/10.1016/B978-0-12-394592-1.00010-7 162 Camille Boutin et al. Cadherin EGF Laminin G HRM GPS Figure 7.1 Schematic representation of the Celsr1 protein. The extracellular domain in- cludes nine cadherin repeats, six epidermal growth factor (EGF)-like domains, two lam- inin G repeats, one hormone receptor motif (HRM), and a G-protein-coupled receptor proteolytic site (GPS). In contrast to typical cadherins which are single-pass proteins, Celsr1–3 cadherins are anchored to the plasma membrane by seven transmembrane domains. The cytoplasmic tail varies in size and is poorly conserved among the three Celsrs. These receptors are thought to be natural chimeras of cell adhesion proteins and signaling receptors, able to convert cell–cell communication cues into intracellular signals. Studies of the latrotoxin receptor latrophilin, a member of LNB–GPCR, showed that cleavage at the GPS occurs intracellularly. However, the N-terminal (cell adhesion domain) and C-terminal (GPCR) fragments remain associated by noncovalent bonds at the plasma membrane (Krasnoperov et al., 2002; Volynski et al., 2004). It is not clear yet whether cleavage at the GPS occurs in Celsr1–3 proteins (see below). In addition to cadherin and EGF-like repeats which confer adhesive properties and facilitate cell–cell communication, the extracellular domain of Celsr1–3 Celsr1–3 Cadherins in PCP and Brain Development 163 contains a HRM which may bind putative ligands, although none has been identified thus far. Celsr genes were first identified in mice (Hadjantonakis et al., 1997). They form a highly conserved family with homologs in ascidians, worms, flies, and vertebrates. The fruit fly Celsr ortholog was cloned independently by two groups and is known as Flamingo (Fmi)orStarry night (Stan)(Chae et al., 1999; Usui et al., 1999). Genetic studies have established a critical role for Fmi/Stan in planar cell polarity (PCP) events, particularly in the stereotypic organization of wing hairs (trichomes) and sensory bristles (Chae et al., 1999; Usui et al., 1999), the alignment of ommatidia in the eye (Mlodzik, 1999), and the asymmetric division of sensory organ precursors (Bellaiche, Gho, Kaltschmidt, Brand, & Schweisguth, 2001; Segalen & Bellaiche, 2009). In these processes, Fmi/Stan interacts genetically and functionally with so-called core PCP proteins. Drosophila core PCP proteins include serpentine receptors Frizzled (fz and fz2 are partially redundant in PCP); the tetraspannin Van Gogh (vang, also named strabismus); and the three cytoplasmic proteins Disheveled (dsh), Prickle (pk), and Diego (dgo) (Adler, 2002; Feiguin, Hannus, Mlodzik, & Eaton, 2001; Gubb et al., 1999; Strutt, Johnson, Cooper, & Bray, 2002; Taylor, Abramova, Charlton, & Adler, 1998; Wolff & Rubin, 1998). In the insect wing, the distribution of core PCP proteins is tightly and dynamically regulated. Initially distributed in all apical junctions, they adopt a transient polarized partition along the proximal–distal axis shortly before the growth of wing hairs, and this partition is essential to the establishment of polarity and the proper orientation of hairs. fmi/stan, fzd, dsh, and dgo are enriched at the distal border of wing cells, whereas fmi/stan, vang, and pk are enriched at the proximal border (Bastock, Strutt, & Strutt, 2003; Jenny, Darken, Wilson, & Mlodzik, 2003; Usui et al., 1999). Given that fmi/stan localizes to both proximal and distal junctions, it was considered a permissive molecule with no active effect on polarity: fmi/stan homodimers would act as a scaffold, promoting cell adhesion and bridging the distal (fz-expressing) side of a cell and the proximal (vang-expressing) side of the adjacent cell. However, more recent data provide evidence that fmi/stan plays an instructive role: its central portion containing the HRM and TM domains interacts physically with fz. Furthermore, fmi/stan selectively recruits fz and vang to opposite cell boundaries, thereby initiating bidirectional polarity signals (Chen et al., 2008). In line with this, genetic analyses in flies demonstrated a mutual requirement for fmi/stan, fz, and dsh to achieve a 164 Camille Boutin et al. correct partition of polarity complexes (Das, Reynolds-Kenneally, & Mlodzik, 2002). In zebrafish, a serine acidic amino acid-rich domain (SE/D) in the CT of Celsr is required for membrane localization of the Frizzled–Disheveled complex, PCP signaling, and convergent extension (Carreira-Barbosa et al., 2009). 1. CELSR1–3 EXPRESSION PATTERNS In mammals, the Celsr family is composed of three members, whereas birds apparently lack Celsr2 (Formstone, 2010) and fish have two, Celsr1a and 1b (Wada, Tanaka, Nakayama, Iwasaki, & Okamoto, 2006). In the mouse, Celsr1–3 genes have similar genomic organizations, with 35 (Celsr1 & Celsr3) or 34 exons (Celsr2). Apart from 30 alternative exons in Celsr2, alternative splicing has not been described. Celsr1–3 expression is regulated spatially and temporally, indicating that they are important for de- velopment. A striking feature is the complementary pattern of Celsr1 and Celsr3 expressions in different developing systems (Formstone & Little, 2001; Shima et al., 2002; Tissir, De Backer, Goffinet, & Lambert de Rouvroit, 2002). In the nervous system, Celsr1 mRNA is heavily expressed in zones of neural stem cell (NSC) proliferation, namely, all ventricular zones during embryonic and early postnatal development, and telencephalic ependymal zones as well as subgranular layer of the dentate gyrus in the mature brain. In contrast, Celsr3 mRNA is absent from NSC and associated with most postmitotic neural cells, whereas Celsr2 mRNA is found in both NSC and postmitotic cells. Expression of Celsr1 abates during early postnatal development, in parallel to decreasing numbers of NSC. That of Celsr3 is downregulated postnatally but persists in the cerebellar granular layer, the hilus of the dentate gyrus, the rostral migratory stream, and the central region of the olfactory bulb. By contrast, Celsr2 expression remains stable throughout life. These expression patterns hint to functions of Celsr1 in NSC, Celsr3 in neural cell maturation, and Celsr2 in development and maintenance of the nervous system. Celsr1–3 mRNAs are also variably expressed in nonneural tissue, such as the skin, lungs, kidney, and digestive and reproductive systems. In the rodent testis, the spatiotemporal pattern of Celsr1–3 expression is somewhat reminiscent of that in the brain. Celsr3 is expressed exclusively in postmeiotic germ cells, and Celsr1 and Celsr2 in Sertoli cells, with a postnatal downregulation of Celsr1 and persistence of Celsr1–3 Cadherins in PCP and Brain Development 165 Celsr2 in the adult (Beall, Boekelheide, & Johnson, 2005; Hadjantonakis, Formstone, & Little, 1998; Johnson, Patel, & Boekelheide, 2000). Polyclonal antibodies against the extra- and intracellular segments of Celsr1 allowed detection of two forms, full length (p400) and cleaved (p85 kD), generated by as yet unidentified proteolysis events not involving the GPS (Formstone, Moxon, Murdoch, Little, & Mason, 2010). In the hindbrain and spinal cord, Celsr1 protein immunoreactivity was detected in the floor and roof plates, as well as in radial neuroepithelial progenitors, not only in the apical but also in their basolateral domain, particularly in end- feet abutting the pial surface (Formstone et al., 2010). In the
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